Paracrine factors of vascular endothelial cells facilitate cardiomyocyte differentiation of mouse embryonic stem cells.
For myocardial regeneration therapy, the low differentiation capability of functional cardiomyocytes sufficient to replace the damaged myocardial tissue is one of the major difficulties. Using Nkx2.5-GFP knock-in ES cells, we show a new efficient method to obtain cardiomyocytes from embryonic stem (ES) cells. The proportion of GFP-positive cells was significantly increased when ES cells were cultured with a conditioned medium from aortic endothelial cells (ECs), accompanied by upregulation of cardiac-specific genes as well as other mesodermal genes. The promotion was more prominent when EC-conditioned medium was added at an early stage of ES cell differentiation culture (Day 0-3). Inhibitors of bone morphogenic protein (BMP), cyclooxygenase (COX), and nitric oxide synthetase (NO) prevented the promotion of cardiomyogenesis by EC-conditioned medium. These results suggest that supplementation of EC-conditioned medium enables cardiomyocytes to be obtained efficiently through promotion of mesoderm induction, which is regulated by BMP, COX, and NOS.
Kado
Miwako
M
Department of Cardiothoracic Surgery, Nagoya University, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan.
Lee
Jong-Kook
JK
Department of Cardiovascular Research, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi 464-8601, Japan. Electronic address: jlee@riem.nagoya-u.ac.jp.
Hidaka
Kyoko
K
Department of Bioscience, Research Institute, National Cardiovascular Center, 5-7-1 Fujishiro-dai, Suita, 565-8565, Japan.
Miwa
Keiko
K
Department of Cardiovascular Research, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi 464-8601, Japan.
Murohara
Toyoaki
T
Department of Cardiology, Graduate School of Medicine, Nagoya University, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan.
Kasai
Kenji
K
Department of Pathology, Aichi Medical University, Nagakute-cho, Aichi, 480-1195, Japan.
Saga
Shinsuke
S
Department of Pathology, Aichi Medical University, Nagakute-cho, Aichi, 480-1195, Japan.
Morisaki
Takayuki
T
Department of Bioscience, Research Institute, National Cardiovascular Center, 5-7-1 Fujishiro-dai, Suita, 565-8565, Japan.
Ueda
Yuichi
Y
Department of Cardiothoracic Surgery, Nagoya University, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan.
Kodama
Itsuo
I
Department of Cardiovascular Research, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi 464-8601, Japan.
D047228:Angiotensin II Type 1 Receptor Blockers / Q000494:pharmacology; D000818:Animals; D001011:Aorta / Q000166:cytology; D019485:Bone Morphogenetic Proteins / Q000037:antagonists & inhibitors; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000378:metabolism / Q000494:pharmacology; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D019332:Endothelin-1 / Q000037:antagonists & inhibitors; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D055879:Gene Knock-In Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000235:genetics; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D011451:Prostaglandin-Endoperoxide Synthases / Q000187:drug effects; D044140:Receptor, Angiotensin, Type 1 / Q000187:drug effects; D014157:Transcription Factors / Q000235:genetics
D047228:Angiotensin II Type 1 Receptor Blockers; D019485:Bone Morphogenetic Proteins; D017077:Culture Media, Conditioned; D016861:Cyclooxygenase Inhibitors; D019332:Endothelin-1; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; C484469:Nkx2-5 protein, mouse; D044140:Receptor, Angiotensin, Type 1; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D019001:Nitric Oxide Synthase; D011451:Prostaglandin-Endoperoxide Synthases